GENE DOPING IN SPORT – PERSPECTIVES AND RISKS

In the past few years considerable progress regarding the knowledge of the human genome map has been achieved. As a result, attempts to use gene therapy in patients’ management are more and more often undertaken. The aim of gene therapy is to replace defective genes in vivo and/or to promote the long-term endogenous synthesis of deficient protein. In vitro studies improve the production of human recombinant proteins, such as insulin (INS), growth hormone (GH), insulin-like growth factor-1 (IGF-1) and erythropoietin (EPO), which could have therapeutic application. Unfortunately, genetic methods developed for therapeutic purposes are increasingly being used in competitive sports. Some new substances (e.g., antibodies against myostatin or myostatin blockers) might be used in gene doping in athletes. The use of these substances may cause an increase of body weight and muscle mass and a significant improvement of muscle strength. Although it is proven that uncontrolled manipulation of genetic material and/or the introduction of recombinant proteins may be associated with health risks, athletes are increasingly turning to banned gene doping. At the same time, anti-doping research is undertaken in many laboratories around the world to try to develop and refine ever newer techniques for gene doping detection in sport. Thanks to the World Anti-Doping Agency (WADA) and other sports organizations there is a hope for real protection of athletes from adverse health effects of gene doping, which at the same time gives a chance to sustain the idea of fair play in sport.     

GENES IN SPORT AND DOPING

Genes control biological processes such as muscle production of energy, mitochondria biogenesis, bone formation, erythropoiesis, angiogenesis, vasodilation, neurogenesis, etc. DNA profiling for athletes reveals genetic variations that may be associated with endurance ability, muscle performance and power exercise, tendon susceptibility to injuries and psychological aptitude. Already, over 200 genes relating to physical performance have been identified by several research groups. Athletes’ genotyping is developing as a tool for the formulation of personalized training and nutritional programmes to optimize sport training as well as for the prediction of exercise-related injuries. On the other hand, development of molecular technology and gene therapy creates a risk of non-therapeutic use of cells, genes and genetic elements to improve athletic performance. Therefore, the World Anti-Doping Agency decided to include prohibition of gene doping within their World Anti-Doping Code in 2003. In this review article, we will provide a current overview of genes for use in athletes’ genotyping and gene doping possibilities, including their development and detection techniques.     

运动能力相关基因的先天遗传与后天转移

对人体生理特性的研究结果显示,部分运动相关基因如α-肌动蛋白-3、血管紧张素I转换酶、Ⅱ型活化素受体B的基因多态性会明显影响运动员的运动天赋和体能。建立优秀运动员基因库,发现和鉴定可影响运动能力的基因变异体,使得在儿童中开展DNA测试,挑选适合某种特殊体育项目的运动天才和优化训练方法具有一定现实操作意义。另一方面,随着滥用基因技术以提高运动能力的可能性不断提高,部分基因有可能作为基因兴奋剂,通过基因转移的方法导入人体,其所涉及的伦理问题、对人类健康及社会的潜在危害等,已经引起了来自自然科学和社会科学不同领域的广泛关注。     

Physical enhancement of human performance: Is law keeping pace with science?

In the area of genometry—the nascent field of science and technology that proposes to apply enhanced understanding of the human genetic code to reshaping our individual and collective destinies—no topic has generated more interest among the general public, as well as in the athletic community, than the potential for physical enhancement of the human body and its performance. Genometric experiments have produced physically enhanced mice, and the production of similarly enhanced humans may not be far off. Although it is not the objective of most genometric research, the day will come when gene-based “treatments” will enable individuals to build muscle or increase endurance faster than is possible through conventional methods. This article describes developments in the area of physical enhancement that may find application in the “gene doping” of athletes. For example, human performance-related genes may be delivered to athletes using tools developed for research in gene therapy; the protein products of these genes may be administered in recombinant form; and recently discovered small-molecule activators of the major genetic regulatory pathways of physical prowess may be taken orally, providing “exercise in a pill”. This article also describes US and international attempts to regulate and punish the use of prohibited techniques for performance enhancement among athletes. As science advances, defining and detecting “gene doping” becomes increasingly complex. Thus, the study of physical enhancement provides an ideal starting point for the interdisciplinary Redefined Destinies Colloquium's examination of the intersection between law and science.     

Illicit organogenesis: Methods and substances of doping and manipulation

Doping and manipulation are undesirable companions of professional and amateur sport. Numerous adverse analytical findings as well as confessions of athletes have demonstrated the variety of doping agents and methods as well as the inventiveness of cheating sportsmen. Besides ‘conventional’ misuse of drugs such as erythropoietin and insulins, experts fear that therapeutics that are currently undergoing clinical trials might be part of current or future doping regimens, which aim for an increased functionality and performance or organs and tissues. Emerging drugs such as selective androgen receptor modulators (SARMs), hypoxia-inducible factor (HIF) complex stabilizers or modulators of muscle fiber calcium channels are considered relevant for current and future doping controls due to their high potential for misuse in sports.Key words: sport, doping, mass spectrometry, anabolics, insulin, HIF, S107     

Affinity-based biosensors as promising tools for gene doping detection

Innovative bioanalytical approaches can be foreseen as interesting means for solving relevant emerging problems in anti-doping control. Sport authorities fear that the newer form of doping, so-called gene doping, based on a misuse of gene therapy, will be undetectable and thus much less preventable. The World Anti-Doping Agency has already asked scientists to assist in finding ways to prevent and detect this newest kind of doping. In this Opinion article we discuss the main aspects of gene doping, from the putative target analytes to suitable sampling strategies. Moreover, we discuss the potential application of affinity sensing in this field, which so far has been successfully applied to a variety of analytical problems, from clinical diagnostics to food and environmental analysis.     

Incongruence in Doping Related Attitudes,Beliefs and Opinions in the Context of Discordant Behavioural Data: In Which Measure Do We Trust?

Background

Social psychology research on doping and outcome based evaluation of primary anti-doping prevention and intervention programmes have been dominated by self-reports. Having confidence in the validity and reliability of such data is vital.

Methodology/Principal Findings

The sample of 82 athletes from 30 sports (52.4% female, mean age: 21.48±2.86 years) was split into quasi-experimental groups based on i) self-admitted previous experience with prohibited performance enhancing drugs (PED) and ii) the presence of at least one prohibited PED in hair covering up to 6 months prior to data collection. Participants responded to questionnaires assessing a range of social cognitive determinants of doping via self-reports; and completed a modified version of the Brief Implicit Association Test (BIAT) assessing implicit attitudes to doping relative to the acceptable nutritional supplements (NS). Social projection regarding NS was used as control.PEDs were detected in hair samples from 10 athletes (12% prevalence), none of whom admitted doping use. This group of ‘deniers’ was characterised by a dissociation between explicit (verbal declarations) and implicit (BIAT) responding, while convergence was observed in the ‘clean’ athlete group. This dissociation, if replicated, may act as a cognitive marker of the denier group, with promising applications of the combined explicit-implicit cognitive protocol as a proxy in lieu of biochemical detection methods in social science research. Overall, discrepancies in the relationship between declared doping-related opinion and implicit doping attitudes were observed between the groups, with control measures remaining unaffected. Questionnaire responses showed a pattern consistent with self-reported doping use.

Conclusions/Significance

Following our preliminary work, this study provides further evidence that both self-reports on behaviour and social cognitive measures could be affected by some form of response bias. This can question the validity of self-reports, with reliability remaining unaffected. Triangulation of various assessment methods is recommended.     

Illicit organogenesis

Doping and manipulation are undesirable companions of professional and amateur sport. Numerous adverse analytical findings as well as confessions of athletes have demonstrated the variety of doping agents and methods as well as the inventiveness of cheating sportsmen. Besides ‘conventional’ misuse of drugs such as erythropoietin and insulins, experts fear that therapeutics that are currently undergoing clinical trials might be part of current or future doping regimens, which aim for an increased functionality and performance or organs and tissues. Emerging drugs such as selective androgen receptor modulators (SARMs), hypoxia-inducible factor (HIF) complex stabilizers or modulators of muscle fiber calcium channels are considered relevant for current and future doping controls due to their high potential for misuse in sports.     

Doping in sports and its spread to at-risk populations: an international review

Doping is now a global problem that follows international sporting events worldwide. International sports federations, led by the International Olympic Committee, have for the past half century attempted to stop the spread of this problem, with little effect. It was expected that, with educational programs, testing, and supportive medical treatment, this substance-abusing behavior would decrease. Unfortunately, this has not been the case. In fact, new, more powerful and undetectable doping techniques and substances are now abused by professional athletes, while sophisticated networks of distribution have developed. Professional athletes are often the role models of adolescent and young adult populations, who often mimic their behaviors, including the abuse of drugs. This review of doping within international sports is to inform the international psychiatric community and addiction treatment professionals of the historical basis of doping in sport and its spread to vulnerable athletic and non-athletic populations.     

Associations between Physical and Cognitive Doping – A Cross-Sectional Study in 2.997 Triathletes

Purpose

This study assessed, for the first time, prevalence estimates for physical and cognitive doping within a single collective of athletes using the randomized response technique (RRT). Furthermore, associations between the use of legal and freely available substances to improve physical and cognitive performance (enhancement) and illicit or banned substances to improve physical and cognitive performance (doping) were examined.

Methods

An anonymous questionnaire using the unrelated question RRT was used to survey 2,997 recreational triathletes in three sports events (Frankfurt, Regensburg, and Wiesbaden) in Germany. Prior to the survey, statistical power analyses were performed to determine sample size. Logistic regression was used to predict physical and cognitive enhancement and the bootstrap method was used to evaluate differences between the estimated prevalences of physical and cognitive doping.

Results

2,987 questionnaires were returned (99.7%). 12-month prevalences for physical and cognitive doping were 13.0% and 15.1%, respectively. The prevalence estimate for physical doping was significantly higher in athletes who also used physical enhancers, as well as in athletes who took part in the European Championship in Frankfurt compared to those who did not. The prevalence estimate for cognitive doping was significantly higher in athletes who also used physical and cognitive enhancers. Moreover, the use of physical and cognitive enhancers were significantly associated and also the use of physical and cognitive doping.

Discussion

The use of substances to improve physical and cognitive performance was associated on both levels of legality (enhancement vs. doping) suggesting that athletes do not use substances for a specific goal but may have a general propensity to enhance. This finding is important for understanding why people use such substances. Consequently, more effective prevention programs against substance abuse and doping could be developed.     

基因兴奋剂

人类基因组计划的完成为基因治疗提供了依据,同时也给基因兴奋剂的产生创造了条件。本文通过综述基因兴奋剂的概念、产生的分子机制,讨论了可能被用作基因兴奋剂的基因,并从基因、转录和蛋白质三个水平探讨了基因兴奋剂的检测策略。     

State-sponsored research on creatine supplements and blood doping in elite Soviet sport

The former Soviet Union began participating in international sport after World War II and soon achieved a dominant position in the Olympic Games and other competitions. The success of Soviet athletic programs led to charges of unfair practices but, because of secrecy surrounding Soviet research in exercise biochemistry, it has been difficult to substantiate these charges. This article presents previously restricted information regarding the development and use of creatine supplements and blood doping in the USSR. Early work by Olexander Palladin established the role of creatine in muscle function. In the 1970s, Soviet scientists showed that oral creatine supplements improved athletic performance in short, intense activities such as sprints. Subsequent studies in the West substantiated these investigations and have led to the widespread acceptance and use of creatine supplements to enhance muscle function and athletic performance. In addition, however, the Soviet government supported the development of blood doping, which is banned by the International Olympic Committee. Blood doping was pervasive in the USSR in the 1970s and 1980s, and was used by many Soviet athletes in the 1976 and 1980 Olympic Games. Open publication and discussion may help to prevent the abuses that can come from secret scientific research.     

Dietary green and white teas suppress UDP-glucuronosyltransferase UGT2B17 mediated testosterone glucuronidation

The anabolic steroid testosterone can be used by athletes to enhance athletic performance and muscle growth. UDP-glucuronosyltransferase (UGT2B17) is the key enzyme involved in the glucuronidation of testosterone to testosterone glucuronide, which also serves as a marker for the testosterone/epitestosterone (T/E) ratio used to detect testosterone abuse in sport. Inhibitors of testosterone glucuronidation could have an impact on circulating testosterone levels, thus aiding performance, as well as potentially affecting the urinary T/E ratio and therefore masking testosterone abuse. Previous reports have revealed that non-steroidal, anti-inflammatory drugs, diclofenac and ibuprofen, inhibit the UGT2B17 enzyme. The aim of this study is to analyse dietary tea samples for inhibition of testosterone glucuronidation and, where inhibition is present, to identify the active compounds. Analysis of testosterone glucuronidation was conducted by performing UGT2B17 assays with detection of un-glucuronidated testosterone using high performance liquid chromatography. The results from this study showed that testosterone glucuronidation was inhibited by the green and white tea extracts, along with specific catechin compounds, notably: epicatechin, epigallocatechin gallate (EGCG) and catechin gallate. The IC50 inhibition value for EGCG was determined, using a Dixon plot, to be 64 μM, equalling the most active NSAID inhibitor diclofenac. Thus, common foodstuffs and their constituents, for the first time, have been identified as inhibitors of a key enzyme involved in testosterone glucuronidation. Whilst these common compounds are not substrates of the UGT2B17 enzyme, we showed that they inhibit testosterone glucuronidation which may have implications on current doping control in sport.     

Second Chances: Investigating Athletes’ Experiences of Talent Transfer

Talent transfer initiatives seek to transfer talented, mature individuals from one sport to another. Unfortunately talent transfer initiatives seem to lack an evidence-based direction and a rigorous exploration of the mechanisms underpinning the approach. The purpose of this exploratory study was to identify the factors which successfully transferring athletes cite as facilitative of talent transfer. In contrast to the anthropometric and performance variables that underpin current talent transfer initiatives, participants identified a range of psycho-behavioral and environmental factors as key to successful transfer. We argue that further research into the mechanisms of talent transfer is needed in order to provide a strong evidence base for the methodologies employed in these initiatives.     

The Effect of Implicitly Incentivized Faking on Explicit and Implicit Measures of Doping Attitude: When Athletes Want to Pretend an Even More Negative Attitude to Doping

The Implicit Association Test (IAT) aims to measure participants’ automatic evaluation of an attitude object and is useful especially for the measurement of attitudes related to socially sensitive subjects, e.g. doping in sports. Several studies indicate that IAT scores can be faked on instruction. But fully or semi-instructed research scenarios might not properly reflect what happens in more realistic situations, when participants secretly decide to try faking the test. The present study is the first to investigate IAT faking when there is only an implicit incentive to do so. Sixty-five athletes (22.83 years ± 2.45; 25 women) were randomly assigned to an incentive-to-fake condition or a control condition. Participants in the incentive-to-fake condition were manipulated to believe that athletes with lenient doping attitudes would be referred to a tedious 45-minute anti-doping program. Attitudes were measured with the pictorial doping brief IAT (BIAT) and with the Performance Enhancement Attitude Scale (PEAS). A one-way MANOVA revealed significant differences between conditions after the manipulation in PEAS scores, but not in the doping BIAT. In the light of our hypothesis this suggests that participants successfully faked an exceedingly negative attitude to doping when completing the PEAS, but were unsuccessful in doing so on the reaction time-based test. This study assessed BIAT faking in a setting that aimed to resemble a situation in which participants want to hide their attempts to cheat. The two measures of attitude were differentially affected by the implicit incentive. Our findings provide evidence that the pictorial doping BIAT is relatively robust against spontaneous and naïve faking attempts. (B)IATs might be less prone to faking than implied by previous studies.     

Direct DNA transfer using electric discharge particle acceleration (ACCELL™ technology)

Direct DNA transfer methods based on particle bombardment have revolutionized plant genetic engineering. Major agronomic crops previously considered recalcitrant to gene transfer have been engineered using variations of this technology. In many cases variety-independent and efficient transformation methods have been developed enabling application of molecular biology techniques to crop improvement. The focus of this article is the development and performance of electric discharge particle bombardment (ACCELL™) technology. Unique advantages of this methodology compared to alternative propulsion technologies are discussed in terms of the range of species and genotypes that have been engineered, and the high transformation frequencies for major agronomic crops that enabled the technology to move from the R&D phase to commercialization. Creation of transgenic soybeans, cotton, and rice will be used as examples to illustrate the development of variety-independent and efficient gene transfer methods for most of the major agronomic crops. To our knowledge, no other gene transfer method based on particle bombardment has resulted in variety-independent and practical generation of large numbers of independently-derived crop plants. ACCELL™ technology is currently being utilized for the routine transfer of valuable genes into elite germplasm of soybean, cotton, bean, rice, corn, peanut and woody species.     

A Multi-Modal Approach to Assessing Recovery in Youth Athletes Following Concussion

Concussion is one of the most commonly reported injuries amongst children and youth involved in sport participation. Following a concussion, youth can experience a range of short and long term neurobehavioral symptoms (somatic, cognitive and emotional/behavioral) that can have a significant impact on one’s participation in daily activities and pursuits of interest (e.g., school, sports, work, family/social life, etc.). Despite this, there remains a paucity in clinically driven research aimed specifically at exploring concussion within the youth sport population, and more specifically, multi-modal approaches to measuring recovery. This article provides an overview of a novel and multi-modal approach to measuring recovery amongst youth athletes following concussion. The presented approach involves the use of both pre-injury/baseline testing and post-injury/follow-up testing to assess performance across a wide variety of domains (post-concussion symptoms, cognition, balance, strength, agility/motor skills and resting state heart rate variability). The goal of this research is to gain a more objective and accurate understanding of recovery following concussion in youth athletes (ages 10-18 years). Findings from this research can help to inform the development and use of improved approaches to concussion management and rehabilitation specific to the youth sport community.     

Gene transfer into eukaryotic cells using activated polyamidoamine dendrimers

The development of efficient methods to transfer genes into eukaryotic cells is important for molecular biotechnology. A number of different technologies to mediate gene transfer have been developed over the last 35 years, but most have drawbacks such as cytotoxicity, low efficiency and/or restricted applicability. Activated polyamidoamine (PAMAM)-dendrimers provide a new technology for gene transfer that offers significant advantages over classical methods. Reagents based on this technology provide high gene transfer efficiencies, minimal cytotoxicity, and can be used with a broad range of cell types. This technology could also be useful for in vivo gene transfer in gene therapy applications.     

Dietary supplement use among adolescent athletes in central Nebraska and their sources of information

The purpose of this study was to investigate the prevalence of dietary supplement use among adolescent athletes. The project was also directed at identifying the sources these student-athletes used for acquiring information about dietary supplements. One hundred thirty nine high school athletes (99 males; 34 females; mean age = 15.8 +/- 1.19 years) volunteered to participate in this study. A 16-question anonymous survey instrument examined use of dietary supplements, reasons for use, type of sport participation, and sources of information regarding dietary supplements. Of the participants in this study, 22.3% (N = 31) reported currently taking dietary supplements. There was no relationship found between dietary supplement use and age. There were a significantly higher number of males reporting current dietary supplement use. Of those who reported to be currently taking dietary supplements, sports performance (N = 25) was the most reported reason for use. There were no significant differences found in reported dietary supplement use between any of the sports. Of the participants, 38.1% (N = 53) listed their coach as their best source of information on dietary supplements. The results of this study offer the current literature some additional insight into trends in supplement use among high school student athletes. Practical implications suggest that it may be necessary to ensure coaches have sufficient knowledge about dietary supplements so that adolescent athletes are receiving accurate information.